The type V collagen group of molecules consists of the recently described collagenous components present in exocytoskeletal matrices. Recent evidence suggests that alterations in the biosynthesis and/or metabolism of type V molecules occur as a consequence of several complex diseases (e.g. atherosclerosis and certain tumors). As all recognized molecular forms of type V collagen molecules contain the Alpha1(V) chain, the ability to assess changes in the mechanisms that regulate its biosynthesis and information about the primary sequence of this collagen chain may have relevance to our understanding of the molecular basis for these diseases. One approach to obtaining such information is to construct complimentary DNA (cDNA) sequences to the mRNA which codes for the Alpha1(V) procollagen chain and to clone these cDNA sequences in bacteria. These clonded sequences can be used measure changes in gene expression, to derive the sequence of the initially synthesized procollagen chain, and to isolate and study the Alpha1(V) procollagen gene. Previous work by this investigator has indicated that HT1 clone of cultured Chinese hamster lung (CHL) cells is a unique system both for isolating the mRNA that codes for the Alpha1(V) procollagen chain and also for dissecting the mechanisms which regulate its biosynthesis. Therefore, the aims of this proposal are to construct, using CHL mRNA as the template, DNA sequences complementary to the Alpha1(V) mRNA, to clone these sequences in bacteria, and to isolate and characterize those recombinants containing the desired cDNA sequences. These recombinant molecules will then be used to: 1) construct additional overlapping cDNA sequences that will encompass the entire sequence of the hamster Alpha1(V) procollagen mRNA; 2) isolate the genomic sequence which codes for this mRNA to allow its characterization; 3) isolate the human Alpha1(V) procollagen mRNA; and 4) to isolate the human gene which codes for this collagen chain. Furthermore, these recombinant cDNA clones will be sequenced to derive the amino acid sequence of the procollagen chain. Completion of the aims of this proposal will provide the information and experimental tools required for future investigations aimed at dissecting the mechanisms that regulate the biosynthesis of the Alpha1(V) procollagen chain and at determining how these controls are altered as a consequence of disease.